Fig. 4: MLKL-driven necroptosis promotes crosstalk between cancer cells and macrophages, leading to macrophage infiltration and activation.

a t-SNE plots of tumour samples from the C57BL/6 orthotopic model mice were generated via scRNA-seq (n = 3 per group). The mice were euthanized 4 weeks after injection. b t-SNE plots of the 28 identified cluster cell populations obtained from the scRNA-seq data. c The proportions of the 28 identified clusters in each sample. d Eleven cell populations were obtained by manual merging in a t-SNE plot on the basis of corresponding marker gene expression. e The epithelial cells were defined into cancer cells (aneuploid) and pancreatic duct cells (diploid) according to the CNV analysis. f tSNE plots from scRNA-seq analysis of OE-vector tumours and OE-MLKL PDAC tumours in C57BL/6 orthotopic model mice and the proportions of the cell types. Aneuploid, cancer cells. g UMAP plots from scRNA-seq analysis of OE-vector tumours and OE-MLKL PDAC tumours in C57BL/6 orthotopic model mice and the proportions of the cell types. h The proportions of different defined macrophage types in the macrophage population. i The single-cell trajectory of macrophages coloured by original cluster identity and the pseudotime trajectory of macrophages. The expression of H2-Aa was projected onto single-cell trajectories. Gene expression values are scaled and log-normalized. j Tumour-infiltrating immune cell score matrix on the basis of T1M0-PDAC (n = 6) and T1M1-PDAC (n = 6) RNA-seq data generated via the QuanTIseq method. The mean of QuanTIseq score in the two groups were shown in the heatmap and compared with two-sided Mann–Whitney U test. k Workflow of the macrophages used for RNA-seq and heatmap showing the DEGs in macrophages co-cultured with PANC-1-OE-Vector cells and PANC-1-OE-MLKL cells for 72 h, after which RNA-seq was performed (n = 3 for each group). l Volcano plot of the DEGs. m KEGG pathway enrichment analysis of the DEGs. n GO enrichment analysis of the DEGs. k Created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license (https://creativecommons.org/licenses/by-nc-nd/4.0/deed.en). Source data are provided as a Source Data file.